title: Identification of a mutated BHK-21 cell line that became less susceptible to Japanese encephalitis virus infection.
abstract: The pathogenesis of Japanese encephalitis virus (JEV) is not definitely elucidated as the initial interaction between virus and host cell receptors required for JEV infection is not clearly defined yet. Here, in order to discover those membrane proteins that may be involved in JEV attachment to or entry into virus permissive BHK-21 cells, a chemically mutated cell line (designated 3A10-3F) that became less susceptible to JEV infection was preliminarily established and selected by repeated low moi JEV challenges and RT-PCR detection for viral RNA E gene fragment. The susceptibility to JEV of 3A10-3F cells was significantly weakened compared with parental BHK-21 cells, verified by indirect immunofluorescence assay, virus plague formation assay, and flow cytometry. Finally, two-dimensional electrophoresis (2-DE) coupled with LC-MS/MS was utilized to recognize the most differentially expressed proteins from membrane protein extracts of 3A10-3F and BHK-21 cells respectively. The noted discrepancy of membrane proteins included calcium binding proteins (annexin A1, annexin A2), and voltage-dependent anion channels proteins (VDAC 1, VDAC 2), suggesting that these molecules may affect JEV attachment to and/or entry into BHK-21 cells and worthy of further investigation.
authors.affiliation: Department of Microbiology, the Fourth Military Medical University, Xi'an, Shaanxi, 710032, PR China.
authors.forename: Tianbing
authors.initials: T
authors.lastname: Ding
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Wenyu
authors.initials: W
authors.lastname: Ma
authors.forename: Junping
authors.initials: J
authors.lastname: Ren
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004664:Encephalitis Virus, Japanese / Q000276:immunology / Q000502:physiology*; D004672:Encephalitis, Japanese / Q000235:genetics* / Q000276:immunology / Q000821:virology; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D009154:Mutation
chemical_list: D008565:Membrane Proteins